Lapatinib and Obatoclax combination therapy

拉帕替尼和 Obatoclax 联合治疗

• 批准号: 8206853
• 负责人: PAUL DENT
• 金额: $ 33.85万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-07 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206853
• 关键词: Animals; Apoptotic; Autophagocytosis; BCL1 Oncogene; BH3 Domain; Breast Cancer Cell; Breast Carcinoma; Cancer Patient; Caspase; Catalytic Domain; Cells; Cessation of life; Colon Carcinoma; Combined Modality Therapy; Cytosol; Data; Diagnosis; Dose; Drops; Drug Combinations; EGFR gene; ERBB2 gene; Endoplasmic Reticulum; Estrogen Receptors; Evaluation; FDA approved; Family; Family member; Fatty acid glycerol esters; Growth; Growth Factor Receptors; Health Benefit; Human Mammary Carcinoma; Image; Immune; In Vitro; Inbred BALB C Mice; Ionizing radiation; Laboratory Study; Malignant Epithelial Cell; Mammary Neoplasms; Mammary gland; Mediating; Mitochondria; Molecular; Mouse Mammary Tumor Virus; Mus; Mutation; Neoplasm Metastasis; Nude Mice; Outcome; PMAIP1 gene; Patients; Pharmaceutical Preparations; Phase; Phase II Clinical Trials; Phosphotransferases; Protein Family; Public Health; Pump; Radiation; Radiation Tolerance; Radiation-Sensitizing Agents; Recurrence; Resistance; Resistance development; Rodent; Small Interfering RNA; Survival Rate; System; Testing; Toxic effect; Translations; Tumor Cell Line; Vesicle; apoptosis inducing factor; base; calreticulin; cancer cell; cancer therapy; cell killing; cell type; cytochrome c; improved; in vivo; inhibition of autophagy; inhibitor/antagonist; killings; knock-down; lapatinib; malignant breast neoplasm; mitochondrial autophagy; mutant; neoplastic cell; public health relevance; tumor; tumor growth

DESCRIPTION (provided by applicant): Prior studies from this laboratory have demonstrated that resistance to the lethal effects of the FDA approved ERBB1 / ERBB2 inhibitor Lapatinib in carcinoma cells is mediated by increased expression of pro-survival BCL-2 family proteins e.g. MCL-1 and BCL-XL. More recently we have gone on to demonstrate that inhibition of MCL-1 and BCL-XL using molecular approaches or using the BCL-2 family inhibitor Obatoclax (GX15-070), a drug in phase II trials, promotes Lapatinib toxicity and reverts Lapatinib resistance. These studies were performed in vitro using a wide range breast and colon cancer cells; drug combination toxicity did not simplistically correlate with ERBB1/2 over-expression. Toxicity, also, was not suppressed by mutant active K- RAS expression or loss of p53 function in the colon cancer cells. This application has three hypotheses: We hypothesize that Lapatinib + Obatoclax treatment causes a toxic form of mitochondrial autophagy. We hypothesize that Lapatinib + Obatoclax treatment interact in vivo in a greater than additive fashion to promote mammary tumor cell killing; reducing tumor growth and metastatic spread. We hypothesize that (Lapatinib + Obatoclax) toxicity in vivo will be magnified in a greater than additive fashion by exposure of the tumor to low doses of ionizing radiation. Specific Aim 1. Determine the molecular mechanisms by which Obatoclax and Lapatinib interact to cause toxic autophagy. The molecular mechanisms of mitochondrial mediated autophagy are not fully understood. We will determine whether Obatoclax causes rapid autophagic degradation of mitochondria via BAK and NOXA (and their altered association with BCL-2 family proteins), with a rapid drop in cellular ATP levels, rather than degradation of the endoplasmic reticulum. We will assess the association of ATG8 and ATG6 (Beclin1) with mitochondrial e.g. HSP60, and ER markers e.g. calreticulin. Specific Aim 2. Determine using orthotopic systems in athymic and immune competent mice whether Obatoclax and Lapatinib interact to kill tumor cells, suppress tumor growth and prolong animal survival. We propose to use multiple tumor cell types to definitively test whether Lapatinib and Obatoclax interact in vivo to reduce tumor growth. We will use human mammary carcinoma BT474 cells growing in the mammary fat pad of athymic mice. We will grow rodent mammary carcinoma MMTV-HER2 cells and 4T1in the mammary fat pad of immune competent syngeneic FVB/NJ and BALB/c mice, respectively; for 4T1 cells we will determine the impact of drug treatment on metastatic tumor cell spread and growth via bioluminescent imaging. Specific Aim 3. Determine whether (Lapatinib + Obatoclax) therapy acts as a radiosensitizer in vivo. We will determine whether (Lapatinib + Obatoclax) treatment enhances tumor cell radiosensitivity in vitro and in vivo. Based on tumor growth / viability data obtained in Aim 2, one of the tumor cell lines will be selected for in vivo evaluation with (Lapatinib + Obatoclax) treatment and radiation exposure in aim 3. Lapatinib is an FDA approved ERBB1 / ERBB2 inhibitor. Obatoclax is in phase II trials. The successful completion of the studies proposed in the application would rapidly facilitate phase I translation of this combination therapy in breast cancer patients. PUBLIC HEALTH RELEVANCE: Public Health Relevance Statement / Project Narrative. Breast cancer is diagnosed in > 200,000 patients per annum, with ~40,000 deaths / year. Thus, although many breast cancer patients have a good 5 year survival rate there is still a major need to improve patient survival to higher levels. Lapatinib is an FDA approved ERBB1 / ERBB2 inhibitor used to treat breast cancer patients. Obatoclax (GX15-070) is an inhibitor of protective BCL-2 family proteins and is in phase II trials. We have discovered that a combination of Lapatinib + Obatoclax rapidly kills naove and Lapatinib resistant breast cancer cells. De novo or developed resistance to cancer therapies, frequently through over-expression of protective BCL- 2 family proteins, and this is a major impediment in obtaining better patient outcomes. As Lapatinib is already FDA approved, the successful completion of the studies proposed in this application would rapidly facilitate phase I translation of the drug combination in patients with recurrent breast cancer.

描述（由申请方提供）：该实验室的既往研究表明，癌细胞对FDA批准的ERBB1/ERBB2抑制剂拉帕替尼致死作用的耐药性是由促生存BCL-2家族蛋白（例如MCL-1和BCL-XL）表达增加介导的。最近，我们继续证明，使用分子方法或使用BCL-2家族抑制剂Obatoclax（GX 15 - 070）（一种II期试验药物）抑制MCL-1和BCL-XL，可促进拉帕替尼毒性并逆转拉帕替尼耐药性。这些研究是在体外使用广泛的乳腺癌和结肠癌细胞进行的;药物联合毒性与ERBB 1/2过表达并不简单相关。在结肠癌细胞中，突变的活性K-RAS表达或p53功能的丧失也不能抑制毒性。本申请有三个假设：我们假设拉帕替尼+Obatoclax治疗引起线粒体自噬的毒性形式。我们假设拉帕替尼+Obatoclax治疗在体内以大于相加的方式相互作用，以促进乳腺肿瘤细胞杀伤;减少肿瘤生长和转移扩散。我们假设（拉帕替尼+奥巴克拉）体内毒性将通过肿瘤暴露于低剂量电离辐射以大于累加的方式放大。具体目标1。确定Obatoclax和Lapatinib相互作用导致毒性自噬的分子机制。线粒体介导的自噬的分子机制尚未完全了解。我们将确定Obatoclax是否通过BAK和NOXA（及其与BCL-2家族蛋白的改变相关性）引起线粒体的快速自噬降解，细胞ATP水平快速下降，而不是内质网降解。我们将评估ATG8和ATG6（Beclin1）与线粒体（如HSP 60）和ER标志物（如钙网蛋白）的相关性。具体目标2。在无胸腺和免疫活性小鼠中使用原位系统确定Obatoclax和拉帕替尼是否相互作用以杀死肿瘤细胞、抑制肿瘤生长并延长动物存活。我们建议使用多种肿瘤细胞类型来明确测试拉帕替尼和Obatoclax是否在体内相互作用以减少肿瘤生长。我们将使用在无胸腺小鼠的乳腺脂肪垫中生长的人乳腺癌BT474细胞。我们将分别在免疫活性同系FVB/NJ和BALB/c小鼠的乳腺脂肪垫中培养啮齿动物乳腺癌MMTV-HER2细胞和4T1细胞;对于4T1细胞，我们将通过生物发光成像确定药物治疗对转移性肿瘤细胞扩散和生长的影响。具体目标3。确定（拉帕替尼+Obatoclax）治疗是否在体内作为放射增敏剂。我们将确定（拉帕替尼+Obatoclax）治疗是否在体外和体内增强肿瘤细胞的放射敏感性。基于目标2中获得的肿瘤生长/活力数据，将选择其中一种肿瘤细胞系用于目标3中的（拉帕替尼+Obatoclax）治疗和辐射暴露的体内评价。拉帕替尼是FDA批准的ERBB1/ERBB2抑制剂。Obatoclax正在进行II期试验。申请中提出的研究的成功完成将迅速促进这种联合疗法在乳腺癌患者中的I期转化。 公共卫生关系： 公共卫生相关性声明/项目叙述。乳腺癌每年在> 200，000名患者中被诊断出，其中约40，000人死亡/年。因此，尽管许多乳腺癌患者具有良好的5年存活率，但仍存在将患者存活率提高到更高水平的主要需求。拉帕替尼是FDA批准的ERBB1/ERBB2抑制剂，用于治疗乳腺癌患者。Obatoclax（GX 15 - 070）是一种保护性BCL-2家族蛋白的抑制剂，目前正处于II期临床试验中。我们已经发现拉帕替尼+奥巴克拉的组合快速杀死耐naove和拉帕替尼的乳腺癌细胞。新生或发展对癌症治疗的耐药性，通常通过保护性BCL-2家族蛋白的过度表达，这是获得更好患者结局的主要障碍。由于拉帕替尼已经获得FDA批准，因此本申请中提出的研究的成功完成将迅速促进药物组合在复发性乳腺癌患者中的I期转化。